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Sirex woodwasps and their symbionts in Alabama forests
- 1. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Sirex woodwasps and their symbionts in Alabama forests Andrea Cole Wahl Sigma Xi Student Showcase
- 2. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Background: Sirex Wasps • Sirex noctilio is a woodwasp associated with Amylostereum spp. fungi, white rot of wood • Females of all Sirex spp. oviposit mixture of eggs, fungal mycelia, and venom into trees • S. nigricornis native to Alabama • Deladenus siricidicola nematode used as biological control agent Introduction Gerard Trichies NEDPN
- 3. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Host Map and Distribution Introduction Native Introduced At risk of introduction Carnegie et al., 2006 USDA APHIS
- 4. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Trapping Survey Flight Phenology
- 5. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Results Flight Phenology 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 MeanInsectsCollectedPerSite Collection Tuskegee Oakmulgee Ranger District Solon Dixon Mean Sirex nigricornis specimens captured Collections from 9/18/14 to 2/25/16
- 6. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Results Flight Phenology Oakmulgee Ranger District Collections from 3/7/15 to 2/20/16 0 0.05 0.1 0.15 0.2 0.25 0.3 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 MeanInsectsCollectedPerSite Collection T. columba S. nigricornis
- 7. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Results Flight Phenology Collections from 9/18/14 to 1/20/16
- 8. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Woodwasp Dissection Molecular Analyses
- 9. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Methodology Molecular Analyses DNA Extraction: Sirex (legs) Amylostereum (mycangia) Deladenus (Sirex eggs) Polymerase Chain Reaction Sequencing
- 10. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Molecular Analyses Molecular Analyses Aligned sequences Drew Phylogenetic trees
- 11. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Molecular Analyses Amylostereum results A. areolatum A. chailletii
- 12. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences, Auburn University Molecular Analyses Deladenus results D. siricidicola D. proximus
- 13. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Background: Tree Defense Once attacked by S. noctilio, a tree begins to exhibit defensive behavior • Constitutive defense: bark thickness • Induced defense: oleoresins, terpenes Chemicals emitted have different functions within a tree • Stress chemicals: α-Pinene, β-Pinene • Defense chemicals: camphene, myrcene, limonene, phellandrene, 4AA Terpene Study
- 14. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Study Design Atmospheric Trial • Atmospheric chambers (paint cans) • Glass petri dishes, no lids • Examined how vaporized terpenes affect fungal growth Tactile Trial • Fungal isolates in direct contact with tepenes on media • Examined how direct contact of terpenes affects fungal growth Terpene Study
- 15. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Materials and Methods Isolation Inoculation and Growth Measurements Terpene Study
- 16. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Atmospheric Trial Results (-)BPinene (+)APinene (-)APinene (+/-)APinene dH2O BMyrcene (+)Camphene (-)Limonene (+)Limonene APhell 4AA Treatment 0 50 100 150 200 250 Fungalgrowth(%ofcontrol) Brazil Australia South Africa 1 New Zealand South Africa 2 South Africa 3 South Africa 4 * Terpene Study **
- 17. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Atmospheric Trial Results (-)BPinene (+)APinene (-)APinene (+/-)APinene dH2O BMyrcene (+)Camphene (-)Limonene (+)Limonene APhell 4AA Treatment 0 20 40 60 80 100 120 140 Fungalgrowth(%ofcontrol) Czech Republic Unknown France Russia Spain Terpene Study ** *
- 18. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Atmospheric Trial Results Terpene Study B-pinene (+)Apinene (-)Apinene (+)(-)Apinene DH2O BMyrcene +Camphene -Limonene +Limonene APhell 4AA Blank Treatment 0 20 40 60 80 100 120 140 160 Fungalgrowth(%ofcontrol) Alabama 1 Alabama 2 S1 N1 *
- 19. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Tactile Trial Results B-pinene (+)Apinene (-)Apinene (+)(-)Apinene DH2O BMyrcene +Camphene -Limonene +Limonene APhell 4AA Treatment 0 50 100 150 200 250 Fungalgrowth(%ofcontrol) Brazil Australia South Africa 1 New Zealand South Africa 2 South Africa 3 South Africa 4 Terpene Study
- 20. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Tactile Trial Results B-pinene (+)Apinene (-)Apinene (+)(-)Apinene DH2O BMyrcene +Camphene -Limonene +Limonene APhell 4AA Treatment 0 20 40 60 80 100 120 140 160 180 200 Fungalgrowth(%ofcontrol) Czech Republic Unknown France Spain Terpene Study *
- 21. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Tactile Trial Results Terpene Study (-)BPinene (+)APinene (-)APinene (+/-)APinene dH2O BMyrcene (+)Camphene (-)Limonene (+)Limonene APhell 4AA Treatment 0 20 40 60 80 100 120 140 160 180 Fungalgrowth(%ofcontrol) Alabama (1) Alabama (2) S1 N1
- 22. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Potential Competitors Competition Study Fraser McKee Total Captured Insects of Concern 2014 Woodwasps Ambrosia beetles Bark beetles 13.7% 9.7% 76.6%
- 23. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Materials and Methods Competition Study Inoculation and Growth Measurements
- 24. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Amylostereum vs. Leptographium Competition Study A. chailletii L. terebrantis A. areolatum L. terebrantis
- 25. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Overall results Competition Study
- 26. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Overall Conclusions • No invasive S. noctilio were captured through the duration of the surveys • Bark and ambrosia beetle peak flight periods were earlier in the year than S. nigricornis, suggesting they could reduce substrate for S. nigricornis • S. nigricornis was found to carry A. areolatum and D. siricidicola, two species typically associated with S. noctilio. D. siricidicola was found to be non- genera specific as a parasite, leaving questions over whether it could be used as a biological control agent in the United States • Defense chemicals: α-Phellendrene and 4-AA significantly reduced growth rates of hyphae in comparison to a dH2O control. Stress chemicals: α-Pinene and β-Pinene did not have an adverse affect on hyphae growth rates. β- Myrcene affected growth of hyphae differently when in direct or indirect contact • Overall, A. areolatum from abroad was a poor competitor for Leptographium spp., while a native isolate of A. chailletii (15B) outcompeted both Leptographium spp. • If S. noctilio was introduced into Alabama forests, it would likely have enough competition that it would not be an economically damaging pest as it has been in the Southern Hemisphere Summary and Conclusions
- 27. Forest Health Dynamics Laboratory School of Forestry and Wildlife Sciences – Auburn University Acknowledgments Graduate Students Jessica Ahl Gifty Acquah Nick Barnwell Jeff Chieppa Pratima Devkota Shrijana Duwadi Charles Essien John Mensah Adam Trautwig Staff Tessa Bauman Sarah Peaden Dalton Smith Major Professor Dr. Lori Eckhardt Committee Members Dr. Kathy Lawrence Dr. Andrew Liebhold Dr. Bernard Slippers Other Mentors Dr. Ryan Nadel Dr. Scott Enebak Insect Collections US Forest Service Foresters Solon Dixon Center Staff Undergraduate Workers Jordan Heath Cody Hartzog Caleb Killough Ashton Newman Chase Seals Wilson Strickland Nick Yashko Cora Yates FABI Katrin Fitza Izette Greyling Dr. Gudrun Dittrich-Schröder Dr. Irene Barnes
Editor's Notes
- This is part of the ongoing Sirex project- You should put your name on this slide, since you are the one giving the talk. Ask where
- Sirex woodwasps are a major economic pest in the southern hemisphere, causing millions of dollars of damage to commercial forests. Sirex noctilio was introduced into the Northeast US in 2004 in contaminated wood packing material. It has since spread from New York to Pennsylvania, but is not currently found in the Southeast. Sirex is not currently devastating commercial forests in North America as it has in the southern hemisphere.
- Sirex woodwasps are a major economic pest in the southern hemisphere, causing millions of dollars of damage to commercial forests. Sirex noctilio was introduced into the Northeast US in 2004 in contaminated wood packing material. It has since spread from New York to Pennsylvania, but is not currently found in the Southeast. Sirex is not currently devastating commercial forests in North America as it has in the southern hemisphere.
- These three field sites were chosen to set up a latitudinal gradient Solon Dixon Center was chosen because is near port of Mobile, where S. noctilio would likely be introduced (if found, but was not found) Black panel flight intercept traps were used, baited with a mixture of Alpha/ beta Pinene, where insects fly towards the chemical, hit the trap, and fall into collection cup that is filled with a diluted anti-freeze mixture. Specimens were collected every two weeks, brought back to lab, morphologically identified to species.
- Note greater number of females collected in 2014 season Distinct end to collection period both years when a hard freeze hits (End of December both years)
- Note low mean of specimens collected T. Columba seems to emerge earlier in season than S. nigricornis
- Note low mean of specimens collected T. Columba seems to emerge earlier in season than S. nigricornis
- Andrea is currently away working on another aspect of this project, using molecular techniques to identify what species of woodwasps, fungi, and nematodes are in Alabama’s forests
- Andrea is currently away working on another aspect of this project, using molecular techniques to identify what species of woodwasps, fungi, and nematodes are in Alabama’s forests
- Andrea is currently away working on another aspect of this project, using molecular techniques to identify what species of woodwasps, fungi, and nematodes are in Alabama’s forests
- Andrea is currently away working on another aspect of this project, using molecular techniques to identify what species of woodwasps, fungi, and nematodes are in Alabama’s forests
- Andrea is currently away working on another aspect of this project, using molecular techniques to identify what species of woodwasps, fungi, and nematodes are in Alabama’s forests
- External defense- oozing, pitching out Chemical defenses change depending on the situation the tree is in. When trees are healthy, they give off “defense chemicals” which have been shown to be fungitoxic or fungistatic. When trees become stressed, they begin to give off stress chemicals, which unfortunately are the same chemicals that are attractive to bark beetles/ woodwasps (due to coevolution)
- Two different trials were conducted: Atmospheric- Atmospheric chambers were created using standard paint cans. Glass petri dishes were inoculated and randomly stacked in the cans (no lids), 5 mL of the designated terpen was in an uncovered glass petri dish in the bottom of the chamber. The cans were then sealed and allowed to grow for one week, then growth of fungus was traced and surface area was measured This study showed how terpenes affect the mycelia of the fungus in an indirect, volatile setting Tactile- 1 mL of designated chemical was pipetted directly onto plate, allowed to dry, then isolate was placed onto plate. The growth was measured at 3, 5, 7 days after inoculation (same method, surface area). This showed how mycelia were affected when in direct contact with the chemicals
- Isolation- mycelia were grown from native Sirex nigricornis trapped near campus, other isolates from around the world were obtained from FABI Inoculation/ growth- fungal isolates were grown out, then punched and transferred to the new plates in each respective study Measurements- growth of mycelia was traced onto transparencies (3,5,7 days in tactile trial, 7 days vapor trial), where surface area was measured with planimeter
- Green bar shows chemicals that are “stress chemicals” Orange bar shows chemicals that are “defense chemicals” – in the isolates from the southern hemisphere camphene, (-) limonene, and 4AA significantly reduced growth rates of mycelia compared to the control (water)
- Isolates from northern hemisphere (native range) show same results, but note how much slower growing isolates are overall (look at percentages on side of graph)
- Isolates from northern hemisphere (native range) show same results, but note how much slower growing isolates are overall (look at percentages on side of graph)
- Tactile trial results are similar, but chemicals that are significantly less than control are B- myrcene and 4AA Note that the chemicals deemed as defense chems (orange bar) were so noxious that they not only stunted the growth of the mycelia, they melted the plastic petri dishes in some cases.
- Same results isolates from southern hemisphere, just again reiterate that northern hemisphere isolates grow slower (are less virulent) in native range, meaning isolates that have been introduced are more virulent
- Same results isolates from southern hemisphere, just again reiterate that northern hemisphere isolates grow slower (are less virulent) in native range, meaning isolates that have been introduced are more virulent